New Anti-RSV Nucleoprotein Monoclonal Antibody Pairs Discovered Using Rabbit Phage Display Technology

IF 3 Q3 IMMUNOLOGY

Antibodies Pub Date : 2023-11-08 DOI:10.3390/antib12040073

Pierre-Emmanuel Baurand, Jérémy Balland, Emilia Galli, Suvi Eklin, Rémy Bruley, Laurence Ringenbach

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引用次数: 0

Abstract

Human respiratory syncytial virus (hRSV) is one of the major contagious viruses and causes complicated respiratory issues, especially in young children. The sensitive and fast detection of hRSV is critical for taking the most effective actions. In the present study, rabbit antibodies against the hRSV nucleoprotein (NP) were developed using phage display technology. A female rabbit was immunized with an hRSV strain A2 recombinant NP. A Fab library was built and sorted during two successive panning rounds for strain B and the A2 NP (recombinant preparations), respectively. The choice of candidates was performed using ELISA on the two NP strains. The obtained library was 3 × 106 cfu/mL, with an insertion rate of >95%. The two panning rounds permitted an enrichment factor of 100. ELISA screening allowed us to obtain 28 NP-specific Fab candidates. Among them, 10 retained candidates were reformatted into rabbit full IgG; thereafter, pairing tests on the recombinant strains and native lysate samples were performed. After the pairing tests on the recombinant strains, 53 pairs were identified. Eleven pairs were identified as being able to detect RSVs from native lysates. This work presents new high-potential monoclonal antibodies mAbs (mAbs), which would benefit from lateral flow testing data with patient materials.

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利用兔噬菌体展示技术发现新的抗rsv核蛋白单克隆抗体对

人呼吸道合胞病毒(hRSV)是一种主要的传染性病毒，可引起复杂的呼吸道疾病，特别是在幼儿中。灵敏、快速地检测出hRSV对于采取最有效的行动至关重要。利用噬菌体展示技术，制备了兔抗hRSV核蛋白(NP)的抗体。用hRSV A2重组NP株免疫雌性兔。菌株B和A2 NP(重组制剂)分别在连续两轮筛选中建立Fab库并进行分类。对两株NP菌株进行ELISA筛选。得到的文库为3 × 106 cfu/mL，插入率为95%。两轮淘金使浓缩系数达到100。ELISA筛选使我们获得了28个np特异性候选Fab。其中10个保留的候选物重组为兔全IgG;然后，对重组菌株和天然裂解液样品进行配对试验。对重组菌株进行配对试验，鉴定出53对重组菌株。鉴定出11对能够从天然裂解物中检测到rsv。这项工作提出了新的高潜力单克隆抗体mab (mab)，这将受益于患者材料的侧流测试数据。

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来源期刊

Antibodies IMMUNOLOGY-

CiteScore

7.10

自引率

6.40%

发文量

审稿时长

11 weeks

期刊介绍： Antibodies (ISSN 2073-4468), an international, peer-reviewed open access journal which provides an advanced forum for studies related to antibodies and antigens. It publishes reviews, research articles, communications and short notes. Our aim is to encourage scientists to publish their experimental and theoretical results in as much detail as possible. There is no restriction on the length of the papers. Full experimental and/or methodical details must be provided. Electronic files or software regarding the full details of the calculation and experimental procedure - if unable to be published in a normal way - can be deposited as supplementary material. This journal covers all topics related to antibodies and antigens, topics of interest include (but are not limited to): antibody-producing cells (including B cells), antibody structure and function, antibody-antigen interactions, Fc receptors, antibody manufacturing antibody engineering, antibody therapy, immunoassays, antibody diagnosis, tissue antigens, exogenous antigens, endogenous antigens, autoantigens, monoclonal antibodies, natural antibodies, humoral immune responses, immunoregulatory molecules.

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